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A method for releasing tumor cells captured by red blood cell biomimetic materials

A technology of bionic materials and tumor cells, which is applied in the field of releasing tumor cells captured by red blood cell bionic materials, and can solve problems such as red blood cell deformation, ion concentration difference, and red blood cell bursting.

Active Publication Date: 2022-08-05
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are two types of red blood cell lysates on the market, one is enzyme lysate, which contains enzymes that can attack specific red blood cell surface antigens, which can cause red blood cell deformation, expansion of biological channels, expansion and cracking, or cause red blood cell denaturation; the other is In the ammonium chloride lysate, ammonium ions cannot pass through the cell membrane, but other ions can pass through, which causes the difference in ion concentration inside and outside the cell, that is, the osmotic pressure difference, which eventually causes the external water to diffuse into the cell and cause the red blood cell to burst.
These two types of red blood cell lysates are difficult to completely disintegrate the biphospholipid molecular layer of the cell membrane, so the DSPE-PEG-X molecules modified based on the hydrophilic and hydrophobic forces are also difficult to be released by the cell membrane, so the denaturation after being affected by the red blood cell lysate The binding force between red blood cells and circulating tumor cells still exists, so it is difficult to achieve efficient release of circulating tumor cells
In addition, the erythrocyte lysate cannot achieve 100% accurate lysing of red blood cells, and it will always cause more or less lysis of other types of cells

Method used

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  • A method for releasing tumor cells captured by red blood cell biomimetic materials
  • A method for releasing tumor cells captured by red blood cell biomimetic materials
  • A method for releasing tumor cells captured by red blood cell biomimetic materials

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0038] image 3 The preparation of folic acid-modified erythrocytes is as follows: adding DSPE-PEG-FA to the erythrocyte dispersion, after incubation, centrifuging and washing to obtain burr spherical erythrocytes with surface-modified folic acid. Among them, the ratio of primordial red blood cells to DSPE-PEG-FA is per 5 × 10 7 0.02 mg DSPE-PEG-FA was added to each erythrocyte.

[0039] Figure 4 The preparation of DSPE-PEG-FITC modified erythrocytes is the same as figure 2 Spike spherocytes modified with DSPE-PEG-FITC in .

[0040] Figure 5 The preparation of the erythrocytes modified by DSPE-PEG-biotin is as follows: adding DSPE-PEG-biotin to the erythrocyte dispersion liquid, after incubation, centrifuging and washing to obtain echinocytes with surface-modified biotin. Among them, the ratio of primordial red blood cells to DSPE-PEG-biotin is per 5 × 10 7 0.04 mg DSPE-PEG-biotin was added to each erythrocyte.

Embodiment 1

[0041] [Example 1] Hela cells captured by surface-modified erythrocytes with DSPE-PEG-FA were released with DMEM complete medium

[0042] (1) Capture

[0043] The system uses erythrocytes surface-modified with DSPE-PEG-FA to capture Hela cells, and the total volume of the capture system is 0.2 mL.

[0044] The preparation process of erythrocytes with surface-modified DSPE-PEG-FA: adding DSPE-PEG-FA to the erythrocyte dispersion, after incubation, centrifuging and washing to obtain burr spherical erythrocytes with surface-modified folic acid. Among them, the ratio of primordial red blood cells to DSPE-PEG-FA is per 5 × 10 7 0.02 mg DSPE-PEG-FA was added to each erythrocyte.

[0045] Capture Hela Cell Process: Quantitative Take 10 5 HeLa cells were washed and dispersed with PBS, to which 10 7 The red blood cells modified with DSPE-PEG-FA on the surface were mixed evenly, centrifuged at 1500 rpm for 3 min in a centrifuge, and then incubated at 37°C for 1 h. The effect of pure...

Embodiment 2

[0049] [Example 2] MCF-7 cells captured by surface-modified erythrocytes with DSPE-PEG-RGD were released with DMEM high glucose complete medium

[0050] (1) Capture

[0051] The system uses red blood cells modified with DSPE-PEG-RGD on the surface to capture MCF-7 cells, and the total volume of the capture system is 0.4 mL.

[0052] The preparation process of erythrocytes with surface-modified DSPE-PEG-RGD: adding DSPE-PEG-RGD to the erythrocyte dispersion, after incubation, centrifuging and washing to obtain burr spherical erythrocytes with surface-modified DSPE-PEG-RGD. Among them, the ratio between the amount of primitive red blood cells and DSPE-PEG-RGD is every 5 × 10 7 0.02 mg DSPE-PEG-RGD was added to each erythrocyte.

[0053] Capture Hela Cell Process: Quantitative Take 10 5 MCF-7 cells were dispersed and washed with PBS, to which 10 7erythrocytes modified with DSPE-PEG-RGD on the surface, mixed evenly, centrifuged at 1500 rpm in a centrifuge for 3 min, and then i...

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Abstract

The invention discloses a method for releasing tumor cells captured by erythrocyte biomimetic materials, and belongs to the technical field of biomedicine. The present invention finds that DSPE-PEG-X molecules can be detached from the red blood cell membrane through the action of the complete medium. The method of the present invention is to incubate the red blood cell biomimetic material that captures tumor cells with a medium, and then incubate the tumor cells from the red blood cell bionic material. The erythrocyte biomimetic material includes pure erythrocytes modified with DSPE-PEG-X (X represents FA, RGD or biotin), erythrocytes modified by nanomaterials, erythrocyte clusters, plane erythrocyte biomimetic layers, and the like. The tumor cells released by the method of the present invention do not need to be subjected to medium exchange treatment, and can be directly cultured under the condition of complete medium.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for releasing tumor cells captured by erythrocyte biomimetic materials. Background technique [0002] Local or regional recurrence and distant metastasis of tumors pose severe challenges for early diagnosis and cure of tumors. In recent years, molecular biology and clinical studies have shown that circulating tumor cells (CTCs) lead to tumor invasion and the occurrence of micrometastases. CTCs are formed by the original tumor cells shed from the primary tumor and enter the blood system of the human body. CTCs circulate and spread in the peripheral blood, and finally transfer to other tissues and organs of the human body, thereby forming the same type as the primary tumor. of secondary tumors, that is, the metastasis and recurrence of tumors and cancers. By capturing circulating tumor cells in peripheral blood and analyzing the changes in the number of c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2509/00
Inventor 刘威彭伟张陶冶
Owner WUHAN UNIV